This project is directed at understanding, characterizing, and evaluating a novel form of gene product delivery for immunization and treatment: direct DNA injection. Cytokines will be administered intramuscularly to mice in the form of either a cDNA expression vector or preformed protein simultaneously or in sequence with cDNAs for specific antigens from chlamydia in order to elicit a strong immune response. Animals will be followed by evaluation of cytokine response in the circulation, local response to the injected materials at the site of administration, and development and type of immunity. To these ends we have obtained and/or engineered into expressible forms the cDNAs for murine TNF-alpha, murine IL-2, murine IL-4, and murine IL-6. In order to develop the source for the chlamydial antigens needed for these studies, we have generated DNA libraries in a bacterial plasmid system from both Chlamydia trachomatis and Chlamydia pneumoniae. From these libraries we have cloned the genes for the chlamydial major outer membrane proteins and the chlamydial 57kd heat shock protein. These genes have been transferred into bacterial and eukaryotic expression systems.